Rapid Identification of Alkaloids and Flavonoids in Fissistigma oldhamii var. longistipitatum by Ultra High-Performance Liquid Chromatography and Quadrupole Time-of-Flight Tandem Mass Spectrometry.

The herb Fissistigma oldhamii var. longistipitatum has been used for a long time in Asian folk medicine in the treatment of several diseases, including rheumatoid arthritis and other inflammatory conditions. Researchers in China and elsewhere have analyzed and characterized its chemical content. In this study, a UHPLC-Q-TOF-MS/MS method, run in both positive and negative modes, was used to identify the main chemical compounds in dichloromethane extracts of this F. oldhamii variant. A total of 64 compounds, including 44 alkaloids and 20 flavonoids, were rapidly identified or tentatively characterized by comparing the molecular ion peaks and MS2 mass spectrometry fragment ions, combined with the mass spectrometry information of reference substances, appropriate fragmentation ions and related literatures. For the first time, the developed UHPLC-Q-TOF-MS/MS analysis method allows for the determination of 64 compounds from extracts of the F. oldhamii variant. The method presented here produced results that will be useful in further studies of this herb.

New compounds from the stems of Fissistigma acuminatissimum Merr. and their in vitro anti-inflammatory activity.

Three new compounds, 4,5,6,7-tetramethoxy-3-benzoylbenzofuran (1), 4-hydroxy-3,5,6-trimethoxydihydrochalcone-2-O-ß-D-glucopyranoside (2) and 2-hydroxy-3,4,5,6-tetramethoxyphenylethyl benzoate (3) along with five known flavonoids were isolated from the dichloromethane fraction of the stems of Fissistigma acuminatissimum Merr.'s ethanol extracts. The compounds were obtained by chromatographic methods and the structure elucidation was completed primarily on the basis of spectroscopic analyses, all of these compounds were isolated from F. acuminatissimum for the first time. All the fractions and compounds were evaluated for their anti-inflammatory activity against lipopolysaccharide (LPS)-stimulated tumor necrosis factor α (TNF-α) production in RAW264.7 cells in vitro. The dichloromethane fraction showed the most potent inhibition(38.2%) at 60 µg/mL, compound 1 (70.2%) and 3 (65.2%) showed significant inhibition at 10 µM.

Cellular absorption of anthraquinones emodin and chrysophanol in human intestinal Caco-2 cells.

The intestinal absorption characteristics of anthraquinones emodin and chrysophanol were observed by measuring the intracellular accumulation across Caco-2 cells by the reverse-phase high performance liquid chromatography. The intracellular accumulation of chrysophanol was much greater than that of emodin, the maximum absorption of emodin and chrysophanol being 414.02+/-15.28 and 105.56+/-11.57 nmol/l x mg x protein, respectively. The absorption of each anthraquinone was significantly lower at 4 degrees C than that of 37 degrees C. The effects of the transport inhibitors, verapamil, cyclosporine and phloridzin, on the intracellular accumulation were also examined. Verapamil and cyclosporine increased the absorption of emodin and chrysophanol, while phloridzin inhibited their absorption, all in a dose-dependent manner. These results suggest that the absorption characteristics of emodin and chrysophanol were closely related to their special structure with the hydroxy groups. It is also likely that a specific transport system mediated the intracellular accumulation of emodin and chrysophanol across the Caco-2 cells.

[Reparative and therapeutic effect of rhBMP-2m on mice injured by chemotherapy].

AIM: To probe the therapeutic effect of recombinant human bone marrow morphogenetic protein-2 maturation peptide(rhBMP-2m) on mouse bone marrow injury caused by cyclophosphamide (CTX). METHODS: 18 mice were divided randomly into 3 groups, namely CTX injection group(CTX group), BMP therapy group(BMP group) and PBS control group(Control group), 6 mice each group. CTX of 200 mg/kg per mouse was intraperitonealy(IP) injected at a time to BMP group and CTX group so as to establish the experimental model of mouse bone marrow injury. In BMP group, the therapy started from the second day after injection of CTX by using IP injection of 0.5 mg BMP per mouse each day. In control group, PBS was injected only. Changes of peripheral blood leukocyte numbers in 3 groups were observed. On the 5th and 8th day after CTX injection, DNA content in mouse bone marrow karyocytes and variation of cell cycle were analysed by flow cytometry(FCM). Colony forming unit granulocyte-macrophage(CFU-GM) was cultivated simultaneously. RESULTS: On the 4th day after injection of CTX, the leukocyte number in mouse peripheral blood of CTX group dropped to the lowest level, and then picking up gradually. In respect to the variation of the leukocyte number, there was no significant difference between BMP group and CTX group (P>0.05). On the 5th day, the ratio of G(0)/G(1) phase cells in BMP group was notably higher than that of CTX group, and the necrotic and apoptotic rates decreased markedly (P<0.01). On the 8th day, the number of karyocytes in mouse bone marrow of BMP group was obviously more than that of CTX group (P<0.01). CONCLUSION: BMP has some therapeutic effect on mouse bone marrow injury caused by CTX.